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Detection kit for human parvovirus B19 (HPVB19) type IgM (immunoglobulin) antibody and preparation method of detection kit

A human parvovirus and antibody detection technology, applied in the field of immune detection, can solve the problems of cumbersome detection process, long detection time, long incubation time, etc., and achieve the effects of high sensitivity, rapid detection and good specificity

Inactive Publication Date: 2015-12-16
北京中检安泰诊断科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, virus isolation and culture detection requires samples containing live viruses, which increases the possibility of infection
Moreover, it needs to be cultivated in constant temperature equipment; in addition, the cultivation time is long, usually at least 24 hours of cultivation time is required
Electron microscope observation of virus particles requires electron microscope equipment, which has a small application range and troublesome operation. In addition, the selection of observation points may be disturbed by human factors, which may cause missed detection.
Molecular biology detection requires professional equipment and professional laboratory personnel to complete the detection, the detection process is cumbersome, and the detection time is long
Similarly, radioimmunoassay (RIA), enzyme immunoassay (EIA), immunofluorescence (IF), western blotting (WB) and enzyme-linked immunosorbent assay (ELISA) in serological testing require professionals to operate , requires equipment for interpretation, the detection time is long, and it is difficult to promote in clinical application, popularization, on-site detection, etc.

Method used

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  • Detection kit for human parvovirus B19 (HPVB19) type IgM (immunoglobulin) antibody and preparation method of detection kit
  • Detection kit for human parvovirus B19 (HPVB19) type IgM (immunoglobulin) antibody and preparation method of detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] In this embodiment, the human parvovirus B19 type IgM antibody detection kit includes a test strip or a test card, and a sample diluent.

[0037] figure 1 It is a schematic diagram of the structure of the test strip in the human parvovirus B19 type IgM antibody detection kit of the embodiment of the present invention. Such as figure 1 As shown, the test strip includes a reaction plate 1, and a sample pad 2, a gold conjugate pad 3, a reaction membrane 4, and a coarse fiber filter paper 5 adhered to the reaction plate 1.

[0038] Wherein, the gold conjugate pad 3 is coated with a mouse anti-human IgM monoclonal antibody labeled with colloidal gold. The concentration of the mouse anti-human IgM monoclonal antibody is 2.0-10.0 mg / mL. Preferably, the concentration of the mouse anti-human IgM monoclonal antibody is 2 mg / mL. The mouse anti-human IgM monoclonal antibody is a colorless, clear and transparent solution. Measured by polyacrylamide gel electrophoresis (SDS-PAGE), when...

Embodiment 2

[0052] Example 2: Preparation method of human parvovirus B19 type IgM antibody detection kit

[0053] Recombinant HPVB19 antigen coating reaction membrane 4 is used as the detection line; goat anti-mouse IgG polyclonal antibody coating reaction membrane 4 is used as the quality control line; mouse anti-human IgM monoclonal antibody labeled with colloidal gold is coated with gold conjugate pad, assembled Get the test strip.

[0054] (1) Preparation of reaction membrane:

[0055] Dilute the recombinant HPVB19 antigen with the coating diluent to the optimal coating concentration of 1.0mg / mL-1.5mg / mL;

[0056] Dilute the goat anti-mouse IgG polyclonal antibody with coating diluent to the optimal coating concentration of 1.5mg / mL-3.0mg / mL;

[0057] The coating diluent is 0.05M phosphate buffer. The coating diluent is colorless, clear, transparent liquid and no suspended matter. The buffer was diluted twice with purified water, and the pH value was 7.3-7.5 measured with a pH meter. The bu...

Embodiment 3

[0075] Example 3: Detection method of human parvovirus B19 type IgM antibody detection kit

[0076] 1. Test strip test method

[0077] (1) Take out the test strip from the original aluminum foil bag and place it flat on the table;

[0078] (2) Take 20μL of serum or plasma sample, add it to the sample loading point at the lower end of the indicator arrow of the test strip, and add 100μL of sample diluent (about 2-3 drops);

[0079] (3) Interpret the result within 15-20 minutes, and the test result is invalid after 20 minutes.

[0080] (4) Judgment of test results:

[0081] Negative: Only a red band appears at the position of the quality control line.

[0082] Positive: Two red bands appear at the position of the quality control line and the detection line.

[0083] Invalid: There is no red band at the position of the quality control line

[0084] 2. Test card test method

[0085] (1) Take out the test card from the aluminum foil bag, place it on a horizontal work surface, and mark the sample;...

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Abstract

The invention discloses a detection kit for a human parvovirus B19 (HPVB19) type IgM (immunoglobulin) antibody. The detection kit comprises a detection strip, wherein the detection strip comprises a gold conjugate pad and a reaction film, the gold conjugate pad is coated with a colloidal gold labeled mouse-anti-human IgM monoclonal antibody, the reaction film comprises a detection line and a quality control line, the detection line is coated with a recombinant HPVB19 antigen, and the quality control line is coated with a goat-anti-mouse IgG polyclonal antibody. The invention further discloses a preparation method of the kit. The preparation method comprises a reaction film preparation step, a gold conjugate pad preparation step as well as a cutting and assembly step. The kit is simple and convenient to operate, low in cost, good in specificity and high in sensitivity, is suitable for popularization and early-stage diagnosis, can perform single detection and is applicable to epidemiologic investigation and field application.

Description

Technical field [0001] The present invention relates to the field of immune detection, in particular to a detection kit for human parvovirus B19 type IgM antibody and a preparation method thereof. Background technique [0002] Human parvovirus B19 type (HPVB19) is a linear single-stranded DNA virus with a genome size of 5.6 kb, which is classified as the genus Parvovirinae. The virus has strong resistance to external physical effects. The virus particles cannot be inactivated at 56°C for 10 minutes, and lipid solvents have no effect on it. Under the action of formalin, β-propiolactone and γ rays, it may cause damage. Its inactivation. [0003] Human parvovirus B19 is the main source of infection with blood products and children aged 2-7 years. The infections are distributed worldwide. The positive rate of human parvovirus B19 antibody in the population accounts for 60-65%, and there is no significant difference in regional distribution. Human parvovirus B19 infection is generall...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/56983G01N2333/015
Inventor 李海波周晟刘昊
Owner 北京中检安泰诊断科技有限公司
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